| Autor: |
Brothwell JA; Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA., Fortney KR; Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA., Gao H; Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Indiana, USA., Wilson LS; Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, Alabama, USA., Andrews CF; Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, Alabama, USA., Tran TM; Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA.; Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USA.; Ryan White Center for Pediatric Infectious Diseases and Global Health, Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA., Hu X; Division of Pulmonary, Allergy, and Critical Care, Emory University, Atlanta, Georgia, USA., Batteiger TA; Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USA., Barnes S; Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, Alabama, USA.; Targeted Metabolomics and Proteomics Laboratory, University of Alabama at Birmingham, Birmingham, Alabama, USA., Liu Y; Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Indiana, USA.; Department of Biostatistics, Indiana University School of Medicine, Indianapolis, Indiana, USA., Spinola SM; Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA.; Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USA.; Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USA. |
| Abstrakt: |
Few studies have investigated host-bacterial interactions at sites of infection in humans using transcriptomics and metabolomics. Haemophilus ducreyi causes cutaneous ulcers in children and the genital ulcer disease chancroid in adults. We developed a human challenge model in which healthy adult volunteers are infected with H. ducreyi on the upper arm until they develop pustules. Here, we characterized host-pathogen interactions in pustules using transcriptomics and metabolomics and examined interactions between the host transcriptome and metabolome using integrated omics. In a previous pilot study, we determined the human and H. ducreyi transcriptomes and the metabolome of pustule and wounded sites of 4 volunteers (B. Griesenauer, T. M. Tran, K. R. Fortney, D. M. Janowicz, et al., mBio 10:e01193-19, 2019, https://doi.org/10.1128/mBio.01193-19). While we could form provisional transcriptional networks between the host and H. ducreyi, the study was underpowered to integrate the metabolome with the host transcriptome. To better define and integrate the transcriptomes and metabolome, we used samples from both the pilot study ( n = 4) and new volunteers ( n = 8) to identify 5,495 human differentially expressed genes (DEGs), 123 H. ducreyi DEGs, 205 differentially abundant positive ions, and 198 differentially abundant negative ions. We identified 42 positively correlated and 29 negatively correlated human-H. ducreyi transcriptome clusters. In addition, we defined human transcriptome-metabolome networks consisting of 9 total clusters, which highlighted changes in fatty acid metabolism and mitigation of oxidative damage. Taken together, the data suggest a mixed pro- and anti-inflammatory environment and rewired central metabolism in the host that provides a hostile, nutrient-limited environment for H. ducreyi. IMPORTANCE Interactions between the host and bacteria at sites of infection in humans are poorly understood. We inoculated human volunteers on the upper arm with the skin pathogen H. ducreyi or a buffer control and biopsied the resulting infected and sham-inoculated sites. We performed dual transcriptome sequencing (RNA-seq) and metabolic analysis on the biopsy samples. Network analyses between the host and bacterial transcriptomes and the host transcriptome-metabolome network were used to identify molecules that may be important for the virulence of H. ducreyi in the human host. Our results suggest that the pustule is highly oxidative, contains both pro- and anti-inflammatory components, and causes metabolic shifts in the host, to which H. ducreyi adapts to survive. To our knowledge, this is the first study to integrate transcriptomic and metabolomic responses to a single bacterial pathogen in the human host. |
